Biochemical characterization of purified phytase produced from <i>Aspergillus awamori</i> AFE1 associated with the gastrointestinal tract of longhorn beetle (<i>Cerambycidae latreille</i>)

The need for industrially and biotechnologically significant enzymes, such as phytase, is expanding daily as a result of the increased use of these enzymes in a variety of operations, including the manufacture of food, animal feed, and poultry feed. This study sought to characterize purified phytase from <i>A. awamori</i> AFE1 isolated from longhorn beetle for its prospect in industrial applications. Ammonium sulfate precipitation, ion-exchange chromatography, and gel-filtration chromatography were used to purify the crude enzyme obtained from submerged fermentation using phytase-producing media, and its physicochemical characteristics were examined. The homogenous 46.8-kDa phytase showed an 8.1-fold purification and 40.7% recovery. At 70 C and pH 7, the optimum phytase activity was noted. At acidic pH 4–6 and alkaline pH 8–10, it likewise demonstrated relative activity of 88–95% and 67–88%, respectively. It showed 67–70% residual activity between 30 and 70 C after 40 min, and 68–94% residual activity between pH 2 and 12 after 2 h. The presence of Hg⁺, Mg²⁺, and Al³⁺ significantly decreased the enzymatic activity, whereas Ca²⁺ and Cu²⁺ enhanced it. Ascorbic acid increased the activity of the purified enzyme, whereas ethylenediaminetetraacetic acid (EDTA) and mercaptoethanol inhibited it. The calculated values for K_m and V_max were 55.4 mM and1.99 μmol/min/mL respectively. <i>A. awamori</i> phytase, which was isolated from a new source, showed unique and remarkable qualities that may find use in industrial operations such as feed pelleting and food processing.